DNA Origami-Based Multivalent Nanobody Display Platform for Potent Neutralization of Botulinum Neurotoxin Type A.

Area of Science:

• Nanotechnology
• Biochemistry
• Toxicology

Background:

• Botulinum neurotoxins (BoNTs) are highly potent toxins requiring rapid neutralization.
• Nanobodies offer high affinity for toxin neutralization but suffer from short bloodstream half-lives.
• Current anti-BoNT strategies need improvement for efficacy and duration.

Purpose of the Study:

• To develop an advanced anti-BoNT/A system using DNA origami nanostructures to prolong nanobody half-life and enhance neutralization.
• To investigate the efficacy of the DON-ciA-C2 system in a BoNT/A poisoning mouse model.

Main Methods:

• Displaying the BoNT/A-neutralizing nanobody ciA-C2 on DNA origami nanostructures (DONs).
• Creating a multivalent nanoscale surface for enhanced toxin binding and stability.
• Evaluating the DON-ciA-C2 system's efficacy in a mouse model of BoNT/A poisoning.

Main Results:

• The DON-ciA-C2 system demonstrated a prolonged half-life (approx. 32 hours vs. 4 hours for ciA-C2 alone).
• Achieved 100% survival rate in mice challenged with 10 LD50 of BoNT/A under both prophylactic and therapeutic administration.
• The nanostructure acted as a biomimetic trap, enhancing capture, enrichment, and sequestration of BoNT/A.

Conclusions:

• DNA origami nanostructures can significantly enhance the therapeutic potential of nanobodies for treating BoNT poisoning.
• The DON-ciA-C2 system offers a promising, highly efficient neutralization strategy against BoNT/A.
• This approach highlights the potential of DNA origami in precision medicine for protein interaction modulation.